The Role and Expression Regulation of MiR-5047 in the Proliferation and Migration of Breast Cancer Cells

doi:10.13523/j.cb.2012013

China Biotechnology

2021, Vol. 41

Issue (4): 9-17 DOI: 10.13523/j.cb.2012013

The Role and Expression Regulation of MiR-5047 in the Proliferation and Migration of Breast Cancer Cells

LU Yu-xiang,LI Yuan,FANG Dan-dan,WANG Xue-bo,YANG Wan-peng,CHU Yuan-kui(

),YANG Hua(

)

Department of Medical Laboratory, School of Clinical Medicine, Ningxia Medical University, Yinchuan 750004, China

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Abstract

To explore the expression of miR-5047 in breast cancer cells and its role in breast cancer cell proliferation and migration, and to clarify the role of decitabine (DAC) in the regulation of miR-5047 expression. The expression level of miR-5047 in human breast cancer cell lines and normal breast epithelial cells MCF10A was detected by real-time quantitative PCR (qRT-PCR). Transfect miR-5047 mimic and negative control mimic NC into MDA-MB-231 and MCF7 cells, respectively, and verify the transfection efficiency by qRT-PCR. Plate cloning experiments, MTT experiments, and scratch healing experiments were used to detect the proliferation and migration ability of breast cancer cells, and qRT-PCR and Western blot methods were used to detect the expression of related genes and proteins after over-expression of miR-5047. MDA-MB-231 and MCF-7 cells were treated with DAC at final concentrations of 5 μmol/L and 10 μmol/L, and qRT-PCR was used to detect the effect of DAC on miR-5047 expression under different concentrations and treatment time. At the same time, the effect of DAC on the epithelial mesenchymal transition (EMT) of breast cancer cells was detected by morphological observation and Western blot. Compared with normal breast epithelial cells MCF-10A, the expression of miR-5047 in breast cancer cells was significantly down-regulated. Overexpression of miR-5047 can significantly inhibit the proliferation and migration of breast cancer cells, promote the expression of epithelial cell marker E-cadherin, and inhibit the expression of mesenchymal cell marker Vimentin. The expression of miR-5047 can promote the expression of epithelial cell marker E-cadherin and inhibit the expression of mesenchymal cell marker Vimentin. After treating MDA-MB-231 and MCF7 cells with different concentrations of DAC, the expression of miR-5047 was enhanced, and the effect was most significant when 10 μmol/L DAC was used for 48 h. DAC can induce epithelial transformation of MDA-MB-231 cells. The expression of miR-5047 is significantly down-regulated in breast cancer cell lines. Overexpression of miR-5047 can inhibit the proliferation and migration of breast cancer cells. The low expression of miR-5047 in breast cancer cells can inhibit the proliferation and migration of breast cancer cells. DAC treatment can enhance the expression of miR-5047 in breast cancer cells and induce epithelial transformation of the cells.

Key words： MiR-5047 Breast cancer Decitabine (DAC) Cell proliferation/migration Epithelial-mesenchymal transition

Received: 07 December 2020 Published: 30 April 2021

ZTFLH:

Q819

Corresponding Authors: Yuan-kui CHU,Hua YANG E-mail: chuyuankui@163.com;yanghua-126@163.com

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	Yu-xiang LU
	Yuan LI
	Dan-dan FANG
	Xue-bo WANG
	Wan-peng YANG
	Yuan-kui CHU
	Hua YANG

Cite this article:

LU Yu-xiang,LI Yuan,FANG Dan-dan,WANG Xue-bo,YANG Wan-peng,CHU Yuan-kui,YANG Hua. The Role and Expression Regulation of MiR-5047 in the Proliferation and Migration of Breast Cancer Cells. China Biotechnology, 2021, 41(4): 9-17.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2012013 OR https://manu60.magtech.com.cn/biotech/Y2021/V41/I4/9

Table 1 Primer sequences

Fig.1 Relative expression of miR-5047 in normal breast epithelial cells and breast cancer cells ^* P<0.05,^** P<0.01

Fig.2 Measurement of miR-5047 expression in MDA-MB-231 cells and MCF7 cells after transfection of miR-5047 mimics ^*** P<0.001

Fig.3 Effect of miR-5047 overexpression on the proliferation and migration ability of breast cancer cells (a) Clone formation assay (b) MTT assay (c) Scratch healing assay. N.S represents having no significant difference, ^** P<0.01, ^*** P<0.001, ^**** P<0.0001

Fig.4 miR-5047 overexpression induces epithelial mesenchymal transition in breast cancer cells Effects of miR-5047 overexpression on mRNA (a) and protein (b) expression of PCNA, MMP2, Vimentin and E-cadherin in breast cancer cells, ^* P<0.05, ^** P<0.01

Fig. 5 Expression of miR-5047 induced by different concentrations of decitabine at different time points N.S represents having no significant difference; ^* P<0.05, ^** P<0.01, ^*** P<0.001

Fig. 6 DAC induced MET of MDA-MB-231cells (a) Morphological observation of MDA-MB-231cells after 5 μmol/L or 10 μmol/L DAC treatment(magnified 100 times) (b) Detection of cell proliferation, migration and EMT related genes in MDA-MB-231 cells after 5 μmol/L or 10 μmol/L DAC treatment. N.S represents having no significant difference; ^* P<0.05, ^** P<0.01


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